Cai Shengyun, Zhang Pei, Dong Suhe, Li Li, Cai Jianming, Xu Mingjuan
Department of Gynaecology and Obstetrics, Changhai Hospital, Second Military Medical University, Shanghai, China.
Department of Radiation Medicine, Faculty of Naval Medicine, Second Military Medical University, Shanghai, China.
Cell Physiol Biochem. 2018;45(3):1061-1071. doi: 10.1159/000487348. Epub 2018 Feb 7.
BACKGROUND/AIMS: Ovarian cancer (OC) is the fifth leading cause of cancer-related death in women, and it is difficult to diagnose at an early stage. The purpose of this study was to explore the prognostic biological markers of OC.
Univariate Cox regression analysis was used to identify genes related to OC prognosis from the Cancer Genome Atlas(TCGA) database. Immunohistochemistry was used to analyse the level of SPINK13 in OC and normal tissues. Cell proliferation, apoptosis and invasion were performed using MTT assay, flow cytometric analysis and Transwell assay, respectively.
We identified the Kazal-type serine protease inhibitor-13 (SPINK13) gene related to OC prognosis from the Cancer Genome Atlas (TCGA) database by univariate Cox regression analysis. Overexpression of SPINK13 was associated with higher overall survival rate in OC patients. Immunohistochemistry showed that the level of SPINK13 protein was significantly lower in OC tissues than in normal tissues (P < 0.05).In vitro experiments showed that the overexpression of SPINK13 inhibited cellular proliferation and promoted apoptosis. Moreover, SPINK13 inhibited cell migration and epithelial to mesenchymal transition (EMT). SPINK13 was found to inhibit the expression of urokinase-type plasminogen activator (uPA), while recombinant uPA protein could reverse the inhibitory effect of SPINK13 on OC metastasis.
These results indicate that SPINK13 functions as a tumour suppressor. The role of SPINK13 in cellular proliferation, apoptosis and migration is uPA dependent, and SPINK13 may be used as a potential biomarker for diagnosis and targeted therapy in OC.
背景/目的：卵巢癌（OC）是女性癌症相关死亡的第五大主要原因，且早期难以诊断。本研究的目的是探索OC的预后生物学标志物。
采用单变量Cox回归分析从癌症基因组图谱（TCGA）数据库中鉴定与OC预后相关的基因。使用免疫组织化学分析OC组织和正常组织中SPINK13的水平。分别采用MTT法、流式细胞术分析和Transwell法检测细胞增殖、凋亡和侵袭情况。
通过单变量Cox回归分析，我们从癌症基因组图谱（TCGA）数据库中鉴定出与OC预后相关的Kazal型丝氨酸蛋白酶抑制剂-13（SPINK13）基因。SPINK13的过表达与OC患者较高的总生存率相关。免疫组织化学显示，OC组织中SPINK13蛋白水平显著低于正常组织（P < 0.05）。体外实验表明，SPINK13的过表达抑制细胞增殖并促进凋亡。此外，SPINK13抑制细胞迁移和上皮-间质转化（EMT）。发现SPINK13抑制尿激酶型纤溶酶原激活剂（uPA）的表达，而重组uPA蛋白可逆转SPINK13对OC转移的抑制作用。
这些结果表明SPINK13起肿瘤抑制作用。SPINK13在细胞增殖、凋亡和迁移中的作用依赖于uPA，且SPINK13可能作为OC诊断和靶向治疗的潜在生物标志物。
